First record of the myrmicine ant genus Syllophopsis Santschi, 1915 (Hymenoptera: Formicidae) from India with description of a new species.

Here we describe and illustrate Syllophopsis peetersi sp. nov. from Silent Valley National Park, a biodiversity hotspot region of the Western Ghats of India. The discovery also marks a first native report of the genus from the Indian subcontinent. Scanning Electron Microscopy (SEM) analysis was carried out to elucidate the general morphology and sensilla of the new species. The new species is similar to congeners from Madagascar, but with larger differences from species that occur elsewhere.

The basitarsal sulcus gland, a novel exocrine structure in ants.

The basitarsus of the mid- and/or hindlegs of several Amblyoponinae ants shows a deep longitudinal groove or sulcus on its anterior face in workers and queens. Histological examination reveals this sulcus is associated with a conspicuous novel epithelial gland, which brings the number of exocrine glands in the legs of ants to 25. The ultrastructural characteristics of the gland show the presence of a well-developed smooth endoplasmic reticulum. This is indicative for the elaboration of a non-proteinaceous and thus possibly pheromonal secretion. Behavioural observations show that this secretion is collected by the tarsomeres and spread onto the brood and nest, suggesting a role in nestmate recognition. A similar basitarsal sulcus gland was also found in Nothomyrmecia, Paraponera and Tetraponera, which represents both a wide phylogenetic and ecological distribution, as it includes arboreal, ground-dwelling as well as subterranean taxa.

Novel exocrine glands in the foreleg coxae of Discothyrea ants.

Workers, queens and males of all examined Discothyrea species of the 'sauteri group', that have laterally expanded frontal lobes and well-developed antennal scrobes, are characterized by two hairy areas on the outer surface of their procoxae. Histological and ultrastructural examination of Discothyrea sauterirevealed that each of these areas is associated with a novel exocrine gland: the proximal procoxal gland is formed by a cluster of 15 round secretory cells of 34 µm with numerous mitochondria, smooth endoplasmic reticulum and Golgi apparatus.Their ducts have a diameter of 0.5-1 µm.The distal procoxal gland contains 50 secretory cells of 22 µm with numerous vacuoles and lamellar inclusions, and narrow ducts with a diameter of only 0.15-0.2 µm.The differences in ultrastructural appearance and duct diameter indicate that both glands produce a different but probably pheromonal secretion. The function of these novel procoxal glands could not yet be determined, although observation of D.sauteri workers and queens shows that they make frequent and peculiar leg movements, in which the foreleg basitarsus rubs over the coxal hairy areas. The foreleg basitarsus then rubs the ipsilateral hindleg basitarsus and antenna. As a last step of the sequence, the hindleg basitarsus strokes the gaster.In addition to the occurrence of these novel procoxal glands, histological examination of D. sauteri also revealed the presence of yet another novel but smaller procoxal base gland. Ants of the 'testacea group', that have less developed frontal lobes and no antennal scrobes, do not have procoxal hairy areas, although a distinct sculpturation with small pores may occur in the corresponding areas. The related Proceratium japonicum, that has a similar lifestyle as Discothyrea, does not have any of the procoxal glands and does not display the peculiar leg movements as reported for D. sauteri.

Glandular innovations for a tunnelling life: Silk and associated leg glands in Melissotarsus and Rhopalomastix queen and worker ants.

As in other Hymenoptera, adult ants cannot secrete silk, unlike the larvae that spin a cocoon prior to metamorphosis. Fisher and Robertson (1999) first showed the existence of a silk gland in the head of adult Melissotarsus beccarii workers, and we confirm this with detailed histology and ultrastructural comparisons of both queens and workers. This African genus exhibits extreme morphological adaptations (legs, head shape and mandibular muscles) for tunnelling behaviour inside living trees, that underlie an obligate mutualism with scale insects. Rhopalomastix is its sister genus distributed across Asia, and we show that queens and workers also have a silk gland. This lineage of minute workers relies on silk to secure their network of tunnels against other arboreal ants. We show striking differences between these genera in the anatomy and ultrastructure of the cells that secrete silk, especially numerous vacuoles and an unexpectedly branched end apparatus in Melissotarsus. Moreover, the legs of Melissotarsus are much more specialized for tunnelling, and this includes highly expanded basitarsi. The latter house the novel 'Delage-Darchen gland', and we document its anatomy and ultrastructure, suggesting a proteinaceous secretion to harden roofs made of silk combined with wood fragments. The restriction of the Delage-Darchen gland to Melissotarsus, combined with a modified silk gland (an almost three-fold increase in the number of secretory cells, and ultrastructural differences suggestive of higher secretory activity), are evidence of an outstanding evolutionary divergence relative to Rhopalomastix. Synthesis of silk by adults is a significant innovation among ants, but its augmented production in Melissotarsus makes them better adapted for the hazards of arboreal life.

Distal leg structures of the Aculeata (Hymenoptera): A comparative evolutionary study of Sceliphron (Sphecidae) and Formica (Formicidae).

The distal parts of the legs of Sceliphron caementarium (Sphecidae) and Formica rufa (Formicidae) are documented and discussed with respect to phylogenetic and functional aspects. The prolegs of Hymenoptera offer an array of evolutionary novelties, mainly linked with two functional syndromes, walking efficiently on different substrates and cleaning the body surface. The protibial-probasitarsomeral cleaning device is almost always well-developed. A complex evolutionary innovation is a triple set of tarsal and pretarsal attachment devices, including tarsal plantulae, probasitarsomeral spatulate setae, and an arolium with an internal spring-like arcus, a dorsal manubrium, and a ventral planta. The probasitarsal adhesive sole and a complex arolium are almost always preserved, whereas the plantulae are often missing. Sceliphron has retained most hymenopteran ground plan features of the legs, and also Formica, even though the adhesive apparatus of Formicidae shows some modifications, likely linked to ground-oriented habits of most ants. Plantulae are always absent in extant ants, and the arolium is often reduced in size, and sometimes vestigial. The arolium contains resilin in both examined species. Additionally, resilin enriched regions are also present in the antenna cleaners of both species, although they differ in which of the involved structures is more flexible, the calcar in Sceliphron and the basitarsal comb in Formica. Functionally, the hymenopteran distal leg combines (a) interlocking mechanisms (claws, spine-like setae) and (b) adhesion mechanisms (plantulae, arolium). On rough substrate, claws and spine-like setae interlock with asperities and secure a firm grip, whereas the unfolding arolium generates adhesive contact on smooth surfaces. Differences of the folded arolium of Sceliphron and Formica probably correlate with differences in the mechanism of folding/unfolding.

Morphology of the mandibular gland of the ant Paraponera clavata (Hymenoptera: Paraponerinae).

The ant Paraponera clavata (Fabricius, 1775) is the only extant species of Paraponerinae and is widely distributed in Brazilian forests. Aspects of its biology are documented extensively in the literature; however, knowledge of P. clavata internal morphology, specifically of exocrine glands, is restricted to the venom apparatus. The objective of this study was to describe the mandibular gland morphology of P. clavata workers. The mandibular gland is composed of a reservoir connected to a cluster of Type III secretory cells with cytoplasm rich in mitochondria and lipid droplets, similar to that of other ants. Notably, the glandular secretion is rich in protein and has a solid aspect. This is the first morphological description of the mandibular gland of P. clavata. RESEARCH HIGHLIGHTS: This study presents the morphological description of the mandibular gland of Paraponera clavata (Hymenoptera: Paraponerinae). Singular characteristics of the gland are described: the glandular secretion is rich in protein and has a solid aspect.

The cephalic anatomy of workers of the ant species Wasmannia affinis (Formicidae, Hymenoptera, Insecta) and its evolutionary implications.

Despite the ecological significance of ants and the intensive research attention they have received, thorough treatments of the anatomy and functional morphology are still scarce. In this study we document the head morphology of workers of the myrmicine Wasmannia affinis with optical microscopy, µ-computed tomography, scanning electron microscopy, and 3D reconstruction, providing the first complete anatomical treatment of an ant head with a broad array of modern techniques. We discuss the potential of the applied methods to generate detailed and well-documented morphological data sets with increased efficiency. We also address homology problems, particularly in the context of the cephalic digestive tract. According to our analyses the "pharynx" of previous ant studies is homologous to the prepharynx of other insects. We also discuss the phylogenetic potential and functional significance of the observed characters, with internal features such as tentorium and musculature discussed for the first time. Our investigation underlines that detailed anatomical data for Formicidae are still very fragmentary, which in turn limits our understanding of the major design elements underlying the ant bauplan. We attempt to provide a template for further anatomical studies, which will help to understand the evolution of this fascinating group on the phenotypic level.

Morphology and ultrastructure of the infrabuccal pocket and its lining epithelium in workers of Ectomomyrmex javanus (Hymenoptera: Formicidae).

This paper describes the morphology and ultrastructure of the infrabuccal pocket in workers of the carnivorous ant Ectomomyrmex javanus, using SEM and TEM. The infrabuccal pocket is a flexible bag with a diameter of 330 ± 30 µm. In its anterior part, the pocket wall reaches a thickness of 24.45 ± 3.45 µm, which is thicker than the wall lining the rest of the infrabuccal pocket, where it measures 6.87 ± 1.12 µm. The epidermal cells of the wall form a thickened epithelium with a thickness of 10.18 ± 2.50 µm. There are different kinds of hairs inside the pocket, that help in filtering solid pellets. Literature data on the infrabuccal pocket are limited and the function of the thicker epithelium is not yet known. It may provide mechanical strength as the pocket undergoes daily size changes because of the filling up and spitting out of pellets.

Genome Evolution of Bartonellaceae Symbionts of Ants at the Opposite Ends of the Trophic Scale.

Many insects rely on bacterial symbionts to supply essential amino acids and vitamins that are deficient in their diets, but metabolic comparisons of closely related gut bacteria in insects with different dietary preferences have not been performed. Here, we demonstrate that herbivorous ants of the genus Dolichoderus from the Peruvian Amazon host bacteria of the family Bartonellaceae, known for establishing chronic or pathogenic infections in mammals. We detected these bacteria in all studied Dolichoderus species, and found that they reside in the midgut wall, that is, the same location as many previously described nutritional endosymbionts of insects. The genomic analysis of four divergent strains infecting different Dolichoderus species revealed genes encoding pathways for nitrogen recycling and biosynthesis of several vitamins and all essential amino acids. In contrast, several biosynthetic pathways have been lost, whereas genes for the import and conversion of histidine and arginine to glutamine have been retained in the genome of a closely related gut bacterium of the carnivorous ant Harpegnathos saltator. The broad biosynthetic repertoire in Bartonellaceae of herbivorous ants resembled that of gut bacteria of honeybees that likewise feed on carbohydrate-rich diets. Taken together, the broad distribution of Bartonellaceae across Dolichoderus ants, their small genome sizes, the specific location within hosts, and the broad biosynthetic capability suggest that these bacteria are nutritional symbionts in herbivorous ants. The results highlight the important role of the host nutritional biology for the genomic evolution of the gut microbiota-and conversely, the importance of the microbiota for the nutrition of hosts.

Novel thoracic glands in the ant Myopias hollandi.

Besides the common labial and metapleural glands, four novel exocrine glands are described in the thorax of both workers and queens of the ponerine ant Myopias hollandi. From anterior to posterior, these glands were designated as the propleural pit gland, the posterolateral pronotal gland, the anterolateral propodeal gland and the metasternal process gland. They all correspond with class-3 glands, that are made up of bicellular units that each comprise a secretory cell and a duct cell. In the propleural pit gland, the ducts are characterized by a gradually widening diameter, while in the three other glands the ducts show a portion which displays a balloon-like expansion, that on semithin sections stains very dark. For none of these novel glands the function is known as yet, although ultrastructural examination indicates that they produce a non-proteinaceous and therefore possibly pheromonal secretion.

Morphology and ultrastructure of Dufour's and venom glands in the ant Camponotus japonicus Mayr (Hymenoptera: Formicidae).

The Dufour's and venom glands are two important exocrine glands in ants. In this study, the morphology and fine structure of these two glands are described in the ant Camponotus japonicus Mayr. The Dufour glands have a characteristic bilobed shape and show a difference in size and color between the female castes (minor and major workers, alate and dealate queens). The external surface of Dufour's gland shows different features among the female castes. It appears more hypertrophied in major workers than in the other castes, indicating for a more pronounced function. The cells of the glandular epithelium in Dufour's gland are characterized by abundant mitochondria, basal invaginations and developed muscle fibres. The venom gland is morphologically similar in the different female castes, and consists of a venom reservoir, a convoluted gland and a bifurcated free secretory filament, with the convoluted gland appearing as a large cap lying on top of the reservoir. Cells of the convoluted gland and the free secretory filaments contain numerous mitochondria and a prominent end apparatus. The results will contribute to understanding the functional morphology of these glands among the different castes in Camponotus ants.

Morphology and ultrastructure of the mandibular gland in the ant Brachyponera sennaarensis (Hymenoptera, Formicidae).

The 'samsum ant' Brachyponera sennaarensis is an invasive species in Saudi Arabia, where it forms a serious threat because of its painful sting. As part of a morphological survey of the exocrine system of this species, we studied the mandibular gland of males, queens and workers of this species. The gland of males is similar to the common anatomical appearance the mandibular gland has in ants in general, but is considerably different in queens and workers. In both female castes, the secretory cells are grouped in one single cluster, that is surrounded by a thick sheath of connective tissue. The duct cells, that transport the secretion towards the wrinkled reservoir, appear considerably folded. Both the sheath of connective tissue and the folded ducts are considered as a mechanical reinforcement of the gland, although the reason for such reinforcement remains unclear as we are not aware of any peculiar movements of the mandibles in queens and workers. At the ultrastructural level, the secretory cells in all castes are characterized by a well-developed smooth endoplasmic reticulum, which is indicative for the elaboration of a non-proteinaceous and hence possibly pheromonal secretion. The clear structural differences between males and the two female castes, which so far had not been found in other ant species, show that the mandibular gland in B. sennaarensis most likely has a different caste-dependent function.

Titanium and iron titanium oxide nanoparticles in antennae of the migratory ant Pachycondyla marginata: an alternative magnetic sensor for magnetoreception?

The most accepted hypothesis of magnetoreception for social insects is the ferromagnetic hypothesis which assumes the presence of magnetic material as a sensor coupled to sensitive structures that transmit the geomagnetic field information to the nervous system. As magnetite is the most common magnetic material observed in living beings, it has been suggested as basic constituent of the magnetoreception system. Antennae and head have been pointed as possible magnetosensor organs in social insects as ants, bees and termites. Samples of three antenna joints: head-scape, scape-pedicel and pedicel-third segment joints were embedded in epoxi resin, ultrathin sectioned and analyzed by transmission electron microscopy. Selected area electron diffraction patterns and X-ray energy dispersive spectroscopy were obtained to identify the nanoparticle compound. Besides iron oxides, for the first time, nanoparticles containing titanium have been identified surrounded by tissue in the antennae of ants. Given their dimension and related magnetic characteristics, these nanoparticles are discussed as being part of the magnetosensor system.

Imaging with Mass Spectrometry of Bacteria on the Exoskeleton of Fungus-Growing Ants.

Mass spectrometry imaging is a powerful analytical technique for detecting and determining spatial distributions of molecules within a sample. Typically, mass spectrometry imaging is limited to the analysis of thin tissue sections taken from the middle of a sample. In this work, we present a mass spectrometry imaging method for the detection of compounds produced by bacteria on the outside surface of ant exoskeletons in response to pathogen exposure. Fungus-growing ants have a specialized mutualism with Pseudonocardia, a bacterium that lives on the ants' exoskeletons and helps protect their fungal garden food source from harmful pathogens. The developed method allows for visualization of bacterial-derived compounds on the ant exoskeleton. This method demonstrates the capability to detect compounds that are specifically localized to the bacterial patch on ant exoskeletons, shows good reproducibility across individual ants, and achieves accurate mass measurements within 5 ppm error when using a high-resolution, accurate-mass mass spectrometer.

Weaving through a cryptic species: Comparing the Neotropical ants Camponotus senex and Camponotus textor (Hymenoptera: Formicidae).

Camponotus senex (Fr. Smith 1858) and Camponotus textor Forel, 1899 are commonly confused species in the New World tropics. We provide morphological characteristics based on the larvae and adults, behavioural differences, together with evidence from molecular markers (cuticular hydrocarbon profiles, venom differences, nuclear ribosomal ITS-1, and mtDNA COI sequence comparisons) to separate the two species, demonstrating they are not immediately closely related. In conclusion we suggest new reliable morphological characters which can benefit from deeper phenetic analysis, and support the contextual usefulness of non-morphological tools in resolving sibling ant species.

Distribution of the obligate endosymbiont Blochmannia floridanus and expression analysis of putative immune genes in ovaries of the carpenter ant Camponotus floridanus.

The bacterial endosymbiont Blochmannia floridanus of the carpenter ant Camponotus floridanus contributes to its hosts' ontogeny via nutritional upgrading during metamorphosis. This primary endosymbiosis is essential for both partners and vertical transmission of the endosymbionts is guaranteed by bacterial infestation of oocytes. Here we present a detailed analysis of the presence and localisation of B. floridanus in the ants' ovaries obtained by FISH and TEM analyses. The most apical part of the germarium harbouring germ-line stem cells (GSCs) is not infected by the bacteria. The bacteria are detectable for the first time in lower parts of the germarium when cystocytes undergo the 4th and 5th division and B. floridanus infects somatic cells lying under the basal lamina surrounding the ovarioles. With the beginning of cystocyte differentiation, the endosymbionts are exclusively transported from follicle cells into the growing oocytes. This infestation of the oocytes by bacteria very likely involves exocytosis-endocytosis processes between follicle cells and the oocytes. Nurse cells were never found to harbour the endosymbionts. Furthermore we present first gene expression data in C. floridanus ovaries. These data indicate a modulation of immune gene expression which may facilitate tolerance towards the endosymbionts and thus may contribute to their transovarial transmission.

Functional morphology of the maxillary and propharyngeal glands of Monomorium pharaonis (L.).

The maxillary and propharyngeal gland of all 3 castes of Monomorium pharaonis were examined with light and electron microscopy. Although both glands possess a pouch in which secretion can be stored temporarily, a proper reservoir is lacking. The paired maxillary gland opens at the base of the maxilla and consists of 4 secretory cells, which are smaller in workers as compared to queens and males. A digestive role is unlikely as the gland is not directly linked to the digestive system and the amount of rER is negligible. The propharyngeal gland consists of 2 clusters of 16 secretory cells, which open in the pharyngeal atrium through a duct. Secretory cells are smallest in males. Two types of endoplasmic vesicles are observed around the end apparatus, suggesting a release of at least 2 substances. High levels of rER indicate the production of digestive enzymes as one of its functions. No differences between mated versus virgin queens were observed for both glands. Further experiments on chemical and behavioural essays can improve our understanding of the role of both glands in the ant colony. Literature on this topic is very inconsistent. We provide a survey to unravel this chaotic nomenclature issue.

Ultrastructure of the mandibular gland of the ant Myrmoteras iriodum.

The mandibular gland in workers of the formicine ant Myrmoteras iriodum differs from other ants both in its general morphology and ultrastructural organization. The secretory cells appear in a pseudo-epithelial arrangement that gives them a clear polarity. At their apical side, the cells are characterized by a large cup-like extension of the reservoir, from which a bulbous invagination connects to a branched end apparatus. At the basal side, the cells show a labyrinth of basal invaginations, while the lateral cell contacts show clear interdigitations. The cytoplasmic composition reveals the presence of numerous round or elongate inclusions that contain crystalline material. Microtubules are abundant, and locally fibrillar regions are found. The function of the mandibular gland in M. iriodum has not yet been documented, and should be studied using gland extracts and behavioural observations.

Differential scaling within an insect compound eye.

Environmental and genetic influences cause individuals of a species to differ in size. As they do so, organ size and shape are scaled to available resources whilst maintaining function. The scaling of entire organs has been investigated extensively but scaling within organs remains poorly understood. By making use of the structure of the insect compound eye, we show that different regions of an organ can respond differentially to changes in body size. Wood ant (Formica rufa) compound eyes contain facets of different diameters in different regions. When the animal body size changes, lens diameters from different regions can increase or decrease in size either at the same rate (a 'grade' shift) or at different rates (a 'slope' shift). These options are not mutually exclusive, and we demonstrate that both types of scaling apply to different regions of the same eye. This demonstrates that different regions within a single organ can use different rules to govern their scaling, responding differently to their developmental environment. Thus, the control of scaling is more nuanced than previously appreciated, diverse responses occurring even among homologous cells within a single organ. Such fine control provides a rich substrate for the diversification of organ morphology.

Compound eye and ocellar structure for walking and flying modes of locomotion in the Australian ant, Camponotus consobrinus.

Ants are unusual among insects in that individuals of the same species within a single colony have different modes of locomotion and tasks. We know from walking ants that vision plays a significant role in guiding this behaviour, but we know surprisingly little about the potential contribution of visual sensory structures for a flying mode of locomotion. Here we investigate the structure of the compound eye and ocelli in pedestrian workers, alate females and alate males of an Australian ant, Camponotus consobrinus, and discuss the trade-offs involved in optical sensitivity and spatial resolution. Male ants have more but smaller ommatidia and the smallest interommatidial angles, which is most likely an adaptation to visually track individual flying females. Both walking and flying forms of ants have a similar proportion of specialized receptors sensitive to polarized skylight, but the absolute number of these receptors varies, being greatest in males. Ocelli are present only in the flying forms. Each ocellus consists of a bipartite retina with a horizon-facing dorsal retina, which contains retinula cells with long rhabdoms, and a sky-facing ventral retina with shorter rhabdoms. We discuss the implications of these and their potential for sensing the pattern of polarized skylight.